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Infection and Immunity, May 2009, p. 1746-1756, Vol. 77, No. 5
0019-9567/09/$08.00+0     doi:10.1128/IAI.01530-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Differential Expression and Glycosylation of Anaplasma phagocytophilum Major Surface Protein 2 Paralogs during Cultivation in Sialyl Lewis x-Deficient Host Cells{triangledown}

Matthew J. Troese,1,{dagger} Madhubanti Sarkar,2,{dagger},{ddagger} Nathan L. Galloway,1 Rachael J. Thomas,1 Sarah A. Kearns,2 Dexter V. Reneer,2 Tian Yang,3 and Jason A. Carlyon1*

Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, Virginia 23298,1 Department of Microbiology, Immunology, and Molecular Genetics, University of Kentucky College of Medicine, Lexington, Kentucky 40504,2 Paul Lawrence Dunbar High School Math and Science Technology Center, Lexington, Kentucky 405133

Received 17 December 2008/ Returned for modification 13 January 2009/ Accepted 5 February 2009

Many microbial pathogens alter expression and/or posttranslational modifications of their surface proteins in response to dynamics within their host microenvironments to retain optimal interactions with their host cells and/or to evade the humoral immune response. Anaplasma phagocytophilum is an intragranulocytic bacterium that utilizes sialyl Lewis x (sLex)-modified P-selectin glycoprotein ligand 1 as a receptor for infecting myeloid cells. Bacterial populations that do not rely on this receptor can be obtained through cultivation in sLex-defective cell lines. A. phagocytophilum major surface protein 2 [Msp2(P44)] is encoded by members of a paralogous gene family and is speculated to play roles in host adaptation. We assessed the complement of Msp2(P44) paralogs expressed by A. phagocytophilum during infection of sLex-competent HL-60 cells and two HL-60 cell lines defective for sLex expression. Multiple Msp2(P44) and N-terminally truncated 25- to 27-kDa isoforms having various isoelectric points and electrophoretic mobilities were expressed in each cell line. The complement of expressed msp2(p44) paralogs and the glycosyl residues modifying Msp2(P44) varied considerably among bacterial populations recovered from sLex-competent and -deficient host cells. Thus, loss of host cell sLex expression coincided with both differential expression and glycosylation of A. phagocytophilum Msp2(P44). This reinforces the hypothesis that this bacterium is able to generate a large variety of surface-exposed molecules that could provide great antigenic diversity and result in multiple binding properties.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Kontos Medical Sciences Building, 1217 East Marshall Street, Room 215, Richmond, VA 23298-0678. Phone: (804) 628-3382. Fax: (804) 828-9946. E-mail: jacarlyon{at}vcu.edu

{triangledown} Published ahead of print on 17 February 2009.

Editor: R. P. Morrison

{dagger} M.J.T. and M.S. contributed equally to this work.

{ddagger} Present address: Department of Molecular and Cellular Biochemistry, The Ohio State University, 379 Hamilton Hall, 1645 Neil Avenue, Columbus, OH 43210-1218.

Infection and Immunity, May 2009, p. 1746-1756, Vol. 77, No. 5
0019-9567/09/$08.00+0     doi:10.1128/IAI.01530-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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